57 replies to this topic

[proteomist]

In patent application 06127987, Sirtris discusses and shares data on the bioavailability of various resveratrol formulations, including the putative SRT501. I've excerpted several figures from that document below.

The first shows plasma resveratrol concentrations in mice after oral dosing with either 'unprocessed' resveratrol, which has a meam particle size of thirteen microns, or micronized resveratrol (one micron particles) mixed with HPMC (hydroxypropylmethyl cellulose) or HPMC and DOSS (dioctyl sodium sulfosuccinate). In the second part of this figure, they compare oral dosing of the micronized/HPM/DOSS to a TWEEN 80 based preparation. Dosages in all cases are very high.

You can see that the inclusion of excipients, most dramatically here Tween 80, gives a markedly higher peak serum concentration. The Tween 80 peak is quite impressive.

Figure 1


Next let's look at Oral vs IP dosing in mice. You can seen that IP dosing gives much higher peak serum levels.

Figure 2

Now we have some rat data. We're comparing plasma levels for IP versus oral dosing, focusing this time on the Captisol formulation, which is the putative SRT501 based on text in the patent application. Also importantly, we're looking at mostly 50mg/kg doses here instead of 400-600. Again we see that IP administration gives higher concentrations. We also see that the Captisol formulation gives a sharper oral dosage curve, but a lower total area under the curve.

Figure 3

Some thoughts:

Personally, my desired target for serum plasma levels is on the order of 5-10 um. This is what I believe is necessary to get devent Sirt1 activation. If you are a rodent, you can achieve this by oral administration of either ~500mg/kg of raw resvertrol, or something on the order of 100mg/kg or micronized resveratrol prepared with solubilizing agents. Most intriguingly, you may be able to get to these concentrations at a lowly 20mg/kg if the excipient is Tween 80. Thus, a human dose of 1-2 grams in Tween 80 may be quite effective.

Also interesting, we see that an oral dosage of 50mg/kg in rats as the Captisol admixture gives ~6um peak serum levels. This is approximately the dosage used in the SRT501 trials, and this mixture very well may be SRT501 itself, based on some other information in the patent application.

Regarding Tween80, this material is considered consumable and is present in a lot of food preparations. Moreover, it can be obtained fairly cheaply from Sigma or other suppliers. I'm not sure what maximal quantities one can take; at some point it should probably be a laxative. I expect several grams would be safe, however.

I'm going to attempt to make some of these formulations, particularly the Tween form. I'll keep you all posted on how it goes.

Edited by proteomist, 21 May 2007 - 05:49 AM.

[sUper GeNius]

In patent application 06127987, Sirtris discusses and shares data on the bioavailability of various resveratrol formulations, including the putative SRT501. I've excerpted several figures from that document below.

The first shows plasma resveratrol concentrations in mice after oral dosing with either 'unprocessed' resveratrol, which has a meam particle size of thirteen microns, or micronized resveratrol (one micron particles) mixed with HPMC (hydroxypropylmethyl cellulose) or HPMC and DOSS (dioctyl sodium sulfosuccinate). In the second part of this figure, they compare oral dosing of the micronized/HPM/DOSS to a TWEEN 80 based preparation.  Dosages in all cases are very high.

You can see that the inclusion of excipients, most dramatically here Tween 80, gives a markedly higher peak serum concentration. The Tween 80 peak is quite impressive.

Figure 1


Next let's look at Oral vs IP dosing in mice. You can seen that IP dosing gives much higher peak serum levels.

Figure 2

Now we have some rat data. We're comparing plasma levels for IP versus oral dosing, focusing this time on the Captisol formulation, which is the putative SRT501 based on text in the patent application. Also importantly, we're looking at mostly 50mg/kg doses here instead of 400-600. Again we see that IP administration gives higher concentrations. We also see that the Captisol formulation gives a sharper oral dosage curve, but a lower total area under the curve.

Figure 3

Some thoughts:

Personally, my desired target for serum plasma levels is on the order of 5-10 um. This is what I believe is necessary to get devent Sirt1 activation. If you are a rodent, you can achieve this by oral administration of either ~500mg/kg of raw resvertrol, or something on the order of 100mg/kg or micronized resveratrol prepared with solubilizing agents. Most intriguingly, you may be able to get to these concentrations at a lowly 20mg/kg if the excipient is Tween 80. Thus, a human dose of 1-2 grams in Tween 80 may be quite effective.

Also interesting, we see that an oral dosage of 50mg/kg in rats as the Captisol admixture gives ~6um peak serum levels. This is approximately the dosage used in the SRT501 trials, and this mixture very well may be SRT501 itself, based on some other information in the patent application.

Regarding Tween80, this material is considered consumable and is present in a lot of food preparations. Moreover, it can be obtained fairly cheaply from Sigma or other suppliers. I'm not sure what maximal quantities one can take; at some point it should probably be a laxative. I expect several grams would be safe, however.

I'm going to attempt to make some of these formulations, particularly the Tween form. I'll keep you all posted on how it goes.

Thanks for taking a look and providing us with that info.

Is Tween80 any better than something like lecithin or polysorbate80?

[proteomist]

It is polysorbate 80. [thumb]

[sUper GeNius]

It is polysorbate 80. [thumb]

Yeah just discovered that in Wikipedia. Where's a good cheap source of food grade P80?

p.s. Seems like we hit the mother lode with that patent application. P80 is pretty cheap, and if that's all it takes to get decent blood levels of t-res, than we're all in like Flynn...

[rwoodin]

Do we drink it or put it on our head ??

http://www.vitaminwo...tm_nooverride=1

[:o]

[sUper GeNius]

Do we drink it or put it on our head ??

http://www.vitaminwo...tm_nooverride=1

[:o]

I put the stuff on my head way back in '84 when Durk Pearson said it could grow hair. Interesting though, I wonder whether P80 along with t-res could reduce your chance of skin cancer if applied topically.

Also, I guess the t-res suppository thread I started wasn't so outlandish after all, as suppositories are mentioned as a possible delivery method in the patent.

Anyone find a decent source of food grade P80 please post. I'm googling ferociously as we speak...

[proteomist]

I'm working on that too, but so far no luck at consumer scale. I know you can get it from Sigma-Aldrich, but I don't know if they'd sell it to an individual.

It's probably also important to point out that Sirtris used their micronized particles in the Tween formulation. So, bulk coarse resveratrol would work less well. But it would still probably work a whole lot better than without the solubilization.

Anyone find a decent source of food grade P80 please post. I'm googling ferociously as we speak...

[sUper GeNius]

I'm working on that too, but so far no luck at consumer scale. I know you can get it from Sigma-Aldrich, but I don't know if they'd sell it to an individual.

It's probably also important to point out that Sirtris used their micronized particles in the Tween formulation. So, bulk coarse resveratrol would work less well. But it would still probably work a whole lot better than without the solubilization.

Anyone find a decent source of food grade P80 please post. I'm googling ferociously as we speak...

How do we know it works any better than simply adding piperine and quercitin/luteolin?

[sUper GeNius]

I'm working on that too, but so far no luck at consumer scale. I know you can get it from Sigma-Aldrich, but I don't know if they'd sell it to an individual.

It's probably also important to point out that Sirtris used their micronized particles in the Tween formulation. So, bulk coarse resveratrol would work less well. But it would still probably work a whole lot better than without the solubilization.

Anyone find a decent source of food grade P80 please post. I'm googling ferociously as we speak...

How do we know it works any better than simply adding piperine and quercitin/luteolin?

Here ya go. Going on my Visa as we speak!

[URL=http://www.bio-world.com/MoreInfo.asp?EDITEDITEDIT
EDIT.

Shoot, reagent grade, no good.

[Anthony_Loera]

Quite a find proteomist!

Try this link... maybe its the one you are looking for...
http://www.wellnatur...ts/tween80.html


Anthony Loera

[PWAIN]

Do we drink it or put it on our head ??

http://www.vitaminwo...tm_nooverride=1

[:o]

I put the stuff on my head way back in '84 when Durk Pearson said it could grow hair. Interesting though, I wonder whether P80 along with t-res could reduce your chance of skin cancer if applied topically.

Also, I guess the t-res suppository thread I started wasn't so outlandish after all, as suppositories are mentioned as a possible delivery method in the patent.

Anyone find a decent source of food grade P80 please post. I'm googling ferociously as we speak...

What is the situation with using the above linked product? Is it not up to food gradde standards?

[sUper GeNius]

Quite a find proteomist!

Try this link... maybe its the one you are looking for...
http://www.wellnatur...ts/tween80.html


Anthony Loera

Tom found the patent app in this thread, and Proteo' was good enough to read through it for us.

http://www.imminst.o...&f=6&t=16061&s=

Here's what I am wondering. This patent app was dated Dec of 2006. I'm surprised no one has reported it here sooner.

Now, if mixing with P80 is so great, why isn't someone selling t-res that way? Why did not AOR use P80 in their formulation? LEF?

[rwoodin]

Any foods available that are analogous to P80 ?

[proteomist]

Another material commonly used to aid in drug solubilization is PEG (polyethylene glycol), typically an intermediate length polymer like PEG2000 or PEG3000.

The laxative product MiraLax is PEG3350 and can be purchased OTC. I've just found that a ~1:1 mixture of resveratrol and PEG3350 gives much greater suspension of the resveratrol particles in water. While they do not dissolve (and wouldn't be expected to) it keeps them from aggregating into larger clumps. In contrast, in straight water they aggregate very significantly. PEG does this by coating the outside of the resveratrol particles and providing a hydrophilic interface with the water. I've had a solution sitting here for a few hours to see if it would separate again, and so far it still looks just like milk. [thumb]

I don't know how well this will compare with Tween80, but the principle is the same. Interestingly, even on fairly substantial dilution with water after the initial mixing I found the particles remained suspended.

FYI, the recommended laxative dosage is 17 grams dissolved in water, so 2-5 grams isn't a very meaningful dose in that regard.

Edit: On drinking it, the texture is very smooth but the taste is pretty bad. This is a very clean resveratrol product I'm working with, but not pure synthetic. Put directly in the mouth, it has a very slight acrid taste. The taste of the PEG solution is the same, but much more intense. Whether this flavor is resveratrol itself or a minor contaminant, I take the increased flavor to to indicate that the effective solubility is much higher. So, this is a good sign.

It may also help a lot that this particular resveratrol product is very powdery. Very tiny particles. I'll try to measure the particle size under a scope to see how they compare to what Sirtris was using.

[sUper GeNius]

Another material commonly used to aid in drug solubilization is PEG (polyethylene glycol), typically an intermediate length polymer like PEG2000 or PEG3000.

The laxative product MiraLax is PEG3350 and can be purchased OTC. I've just found that a ~1:1 mixture of resveratrol and PEG3350 gives much greater suspension of the resveratrol particles in water. While they do not dissolve (and wouldn't be expected to) it keeps them from aggregating into larger clumps. In contrast, in straight water they aggregate very significantly.  PEG does this by coating the outside of the resveratrol particles and providing a hydrophilic interface with the water. I've had a solution sitting here for a few hours to see if it would separate again, and so far it still looks just like milk. [thumb]

I don't know how well this will compare with Tween80, but the principle is the same. Interestingly, even on fairly substantial dilution with water after the initial mixing I found the particles remained suspended.

FYI, the recommended laxative dosage is 17 grams dissolved in water, so 2-5 grams isn't a very meaningful dose in that regard.

Edit: On drinking it, the texture is very smooth but the taste is pretty bad. This is a very clean resveratrol product I'm working with, but not pure synthetic. Put directly in the mouth, it has a very slight acrid taste. The taste of the PEG solution is the same, but much more intense. Whether this flavor is resveratrol itself or a minor contaminant, I take the increased flavor to to indicate that the effective solubility is much higher. So, this is a good sign.

I just ordered a gallon of Tween80 for 50 bucks. I'll let you know if I actually get it. I'll probably crack open my AOR t-res caps and mix it in some Tween. You used a 1:1 ratio? How much water did you mix in with it?

[sUper GeNius]

Check this out, especially the last paragraph.

http://www.ncbi.nlm....8&dopt=Abstract

[proteomist]

I used about a 1:1 ratio; this is with 98-99% pure resvertrol. Just now I used about three grams t-resv, with about three grams PEG (both weights eyballed, but I'm pretty good at that from my days in a chem lab). Water was to 50 ml, but could have been a lot less. Sirtris used 1/1/3ratio o t-res/Tween80/water if I recall correctly.

[yucca06]

...
Try this link... maybe its the one you are looking for...
http://www.wellnatur...ts/tween80.html


Anthony Loera

You also have this link, if it can help :

http://www.researchl...p?idproduct=158

[yucca06]

I finally found this one :

http://www.thesynerg...plies/sales.htm

...really cheap.

Does it matter if resv 50% is used in the solution, instead of the 98% ?I wonder also if we should add some alcool as a conservative/anti bacterial ? (maybe 1 to 2% benzyl alcool)

Edited by yucca06, 21 May 2007 - 03:01 PM.

[inawe]

I guess all this discussion has to do with bioavailability. In part that's related to particle size. The smaller the particles the better they are absorbed.
Seems that by using a proper solvent one can decrease particle size. I take 13.33 mgr/Kg/day of coarse powder. I know it works because it had a dramatic effect on my lipid profile. I assume that by dissolving it, I transform the coarse powder into smaller particles. The solvent I use is Chianti (I go for the Italian paradox).
So my question is: Would mixing the resveratrol with Tweed 80 be any better than dissolving it in Chianti? If particle size is important, it would be interesting to measure it in both cases.
In the case of Sirtris, maybe they didn't want the mice to get drunk.

[sUper GeNius]

I finally found this one :

http://www.thesynerg...plies/sales.htm

...really cheap.

Does it matter if resv 50% is used in the solution, instead of the 98% ?I wonder also if we should add some alcool as a conservative/anti bacterial ? (maybe 1 to 2% benzyl alcool)

The problem as I see it is that that P80 is sold to be applied topically. What grade is it? It might not be suitable for ingestion.

[malbecman]

Thanks for digging that up and posting those slide, Proteomist. Very interesting results.

I wonder how other food grade emulsifiers beside a detergent might work, eg, lecithin....

[proteomist]

I tried making a solution of Tween 80 and resveratrol in water last night. I used approximately a one to one ratio by mass, and found that the Tween solubilized the resveratrol particles significantly less effectively than does PEG3000. The solution was prone to foaming, and on sitting for a while gave precipitation of some resveratrol particles. Furthermore, it tasted absolutely vile, rather like rancid oil.

I'm going with the PEG myself. Again, this is sold as MiraLax and is tasteless itself.
I don't know that this will accomplish the same high serum levels that Sirtris observed with Tween80. It's possible that Tween80 is doing aditional actions at the gut epithelium that increase resveratrol permeability. However, it's just too nasty for me.

[sUper GeNius]

I tried making a solution of Tween 80 and resveratrol in water last night. I used approximately a one to one ratio by mass, and found that the Tween solubilized the resveratrol particles significantly less effectively than does PEG3000. The solution was prone to foaming, and on sitting for a while gave precipitation of some resveratrol particles. Furthermore, it tasted absolutely vile, rather like rancid oil.

I'm going with the PEG myself. Again, this is sold as MiraLax and is tasteless itself.
I don't know that this will accomplish the same high serum levels that Sirtris observed with Tween80. It's possible that Tween80 is doing aditional actions at the gut epithelium that increase resveratrol permeability. However, it's just too nasty for me.

See my post in the "500mg" thread. Nasty! But what's with the gel? Yuck!

[proteomist]

Probably undispered P80? It takes a lot of mixing to get it to all go in. If you try again, I'd recommed stirring the P80 all the way in, and then adding the resveratrol afterwards.

[sUper GeNius]

Probably undispered P80? It takes a lot of mixing to get it to all go in. If you try again, I'd recommed stirring the P80 all the way in, and then adding the resveratrol afterwards.

Yes, you are correct. I gently heated the water before adding the P80, and then the P80 dissolved quite easily. However, I added another AOR cap, and it leaves an undissolved precipitate at the bottom, no matter how long I stir.

I did read somewhere that Tween 80 can act as an inhibitor of intestinal cyp. Maybe I'll simply use a small bit to wash down the caps.

[lucid]

What order would the effect of a solvent be on bioavailability? Would it be comparable to the bioavailability increase of srt501? Are there any studies done showing that it increases blood plasma levels of tresv when mixed with a solvent?

Since resv is so expensive, increasing bioavailability seems key. Cheers

[maxwatt]

What order would the effect of a solvent be on bioavailability? Would it be comparable to the bioavailability increase of srt501? Are there any studies done showing that it increases blood plasma levels of tresv when mixed with a solvent?

Since resv is so expensive, increasing bioavailability seems key. Cheers

The information is in the patent application referenced at the start of this thread: 200%. PEG 2235 and other PEGs are mentioned as alternative excipients to Tween.

[bugmenot.com]

In patent application 06127987, Sirtris discusses

Where did you find this and what kind of number is that? Searching the uspto database of both applications and issued patents retrieves nothing.

[lucid]

Well I disolved some 98% extract in miralax and it worked really well, no foam or anything at the top. Hopefully I get that 200% increased bioavailability. I can't imagine that the sirtris patent is as simple as that, but I'm not in the mood to read a patent right now And if it is, power to informed consumers!